Editing Synthesis Peptide-Chol

== Synthesis of&nbsp;Cholesterol-acrylate  ==

To a solution of cholesterol (4 g, 10.3 mmol)&nbsp;and TEA (4.3 mL, 31 mmol) in 50 mL of anhydrous THF,&nbsp;acryloyl chloride (1.3 mL, 15.6 mmol) was mixed with 5 mL of&nbsp;anhydrous THF and added dropwise at 0uC under a positive&nbsp;nitrogen flow. Then the mixture was removed from the ice-bath,&nbsp;warmed up to room temperature, and kept stirring overnight.&nbsp;After the removal of the solvent by rotary evaporator, the crude&nbsp;solid was redissolved in 50 mL of CH2Cl2 , and washed with the&nbsp;following solutions: deionized (DI) water, HCl (0.5 M), DI water,&nbsp;NaHCO 3 (1 M), DI water, and brine. The product in CH2Cl2 was&nbsp;dried over anhydrous MgSO4 overnight, and purified by silica gel&nbsp;column chromatography (CH2Cl2 /Hexane 1/1 v/v) after the&nbsp;removal of the solvent by rotary evaporator. Pure product was&nbsp;obtained as a white solid (yield: 35%). 

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== Synthesis of Cholesterol-peptides  ==

Cholesterol-acrylate and peptide (R5H5, R3H3, R5, or R3) conjugation was&nbsp;carried out in a mixture of DMSO and THF at the presence of&nbsp;TEA. The mixture was stirred at 37uC for 2 days under nitrogen&nbsp;protection. Pure product was obtained by dialysis against&nbsp;deionized water using a molecular weight cut-off of 100–500 D&nbsp;dialysis tube from Spectrum Laboratories (Rancho Dominguez,CA, USA) for one day, followed by freeze-drying. 



[[Image:Peptide-chl_conjugate.jpg|600px]]

[[Image:Peptide_chl-conjugate_transfection.jpg]]<br> 

The best one is &nbsp;Ch-R3H3&nbsp;NH2-RRRHHHC-COOH 

[[Category:Transfection]] [[Category:Paul]]